2-methyl-4-sulfonilamido-6-methoxy-3(2h)-pyridazinone for treating coccidiosis



United States Patent 2 METHYL 4 SULFONILAMIDO 6 METHOXY-iaQI-PYRIDAZINONE FOR TREATING COCCID- Miyoshi Ikeda and Fujio Fujioka,Tokyo, and Chutaro Sugiyama and Takenari Nakagome, Nishinomiya, Japan,assignors to Sumitomo Chemical Co., Ltd., Osaka, Japan N0 Drawing. FiledJan. 20, 1966, Ser. No. 521,805

Claims priority, applicationziapan, Jan. 23, 1965,

2 Claims. ci. 167-531) The present invention relates to the use of2-methyl-4- sulfanilamido 6 methoxy-3(2H)-pyridazinone (herein afterreferred to as CS) in the treatment of a poultry coccidiosis.

The instant invention depends upon the fact that CS possesses anunexpectedly remarkable effect against poultry coccidiosis, whereby thedisease can be cured by administering the compound per se or in the formof mixtures with suitable diluents such as poultry feed.

In order to obtain a satisfactory therapeutic effect, the compound CS isadministered for 3 to 4 days at 0.1% (by weight) concentration in feedin the case of ordinary infection, and at 0.2% (by weight) concentrationin the case of severe infection. Alternatively, a 0.1 to 0.4 Weightpercent aqueous solution of CS can be administered for about 3 to 7days.

When the disease occurs outdoors, poultries may be considered to getresistive to the disease by repetition of slight infection. Further, thenumber of oocysts taken at a time may be widely varied in each case andis probably smaller than that experimentally given. Accordingly,complete prevention and treatment of the disease can be safelyaccomplished by consecutive or intermittent administration of arelatively low concentration of the compound CS according to itscharacteristics.

If administered within 4 days after infection by poultry coccidiosis,the compound may be so effective that death of the infected poultriescan be completely prevented and characteristically oocysts are not foundin the feces evacuated by the poultries.

Even when this compound is consecutively administered for 30 days at aconcentration around 0.3%, no side action is found and no substantialdifference is observed in the growth of poultries between theadministered group and the control group. Thus, the compound isextremely low in toxicity and can be used safely.

The result of experimental tests on treatment of artificially infectedpoultries for poultry coccidiosis is set forth below.

(1) Test chickens and method of administration New-born male chickens ofWhite Leghorn were bred in a laboratory for 15 days, from which healthychickens of nearly equal body weight were selected to experiment with ingroups respectively consisting of 10. Each group of the test chickenswas admitted in a wire netting breeding box and was given three times aday paste bait prepared by mixing the drug in a commercially availableformular feed containing no other coccidiostat agent and a half amountof water. The administrations of the drug begin respectively on the 1st2nd, 3rd and 4th day after infection and were continued until thetermination of the test. In view of the characteristics of Eimeriatenella, each test was terminated 9 days after infection.

(2) \Infection material The oocysts of Eimeria tenella were injectedinto the crop of chickens uninfected with coccidiosis. 8 to 9 days afterinfection, the oocysts were collected from the caecum, washed well withwater and allowed to stand in 2.5% potassium bichromate solution at 27to 30 C. for 3 to 4 days. The fresh oocysts were suspended in water tomake a suspension containing the desired numbers (8 X10 or 10 10 ofoocysts per mil. 1 m1. of the suspension was given to each chicken byinjecting directly into the crop.

(3) Judgment of therapeutic eflect The effect of the drug wascollectively judged on the body weight gain, the evacuation ofhemafecia, the activity, the status of intake of feed, the number ofdeaths, the view of post-mortem examination, the detection of oocystsfrom the caecum and feces and the side action of the drug during thetest.

(4) Effect of drug (a) ACTIVITY OF CHICKENS Method of administrationNumber of oocysts infected Slightly dull for 4 to 6 days after the startof the test Considerably dull for 4 to 7 days after the start of thetest. Recovered on the 8th day.

Infected and no administration of t e te Administered everyday with 0.3%concentration. Notddifierent from the uninfected control groupAdministered from the 2nd day with 0.3% con- Consilglerably dull for 4to 7 days after the start Considerably dull for 4 to 7 days after thestart of the test.

centration. Administered everyday with 0.2% concentration. Administeredfrom the 2nd day with 0.2% concentration. Administered from the 3rd daywith 0.2% con- Not different from the uninfected control group.

N 0t 1different from the uninfected control group.

Considerably dull for 4 to 7 days after the start ccntration.

ot the test but better than the infected and non-administered group.

(1)) STATUS OF INTAKE OF FEED Method of administration Number of oocystsinfected Administered from the 3rd day with 0.3% con- Slightly weakappetite. No depression in the centration. amount of feed taken.Administered from the 4th day with 0.3% con- Week appetite for 5 to 6days after the start of eentration. the test Administered everyday with0.1% concentration Weak appetite for 4 to 7 days after the start of1thed 1tesgt. No depression in the amount of ca a en.

Infected and no administration Veg-y lngeak appetite from the 5th day tothe no ay.

Administered everyday with 0.3% concentration.

Administered from the 2nd day with 0.3% concentrntion. Administeredeveryday with 0.2% concentration.

Administered from the 2nd day with 0.2% coneentration.

Administered from the 3rd day with 0.2% concentration.

Slightly week appetite. am ount of feed taken.

No depression in the Extremely weak appetite from the 4th day. Theamount of feed decreased to A for 5 to 9 days after the infection.

Slightly weak appetite. No depression in the anligunt of feed taken.

Weak appetite for 4 to 7 days after the start of the No depression inthe amount of feed en.

THE AMOUNT OF FEED TAKEN BY AVERAGE WEIGHT (G.) PER POULTRY TESTED Daysafter infection of ooeysts Before infection Total amount Age of chickenin days (I)When the number of oocysts infected is 8X10:

0. 3%, everyday.. 21. 0 23. 0 24. 0 25. 0 26. 5 27. 0 27. 0 29. 0 30. 030. 0 262. 5 0.2%, everyday 21. 0 23. 0 24. 0 25. O 26. 5 27. 0 27. 029. 0 30. 0 30.0 262. 5 0.1%, everyday 21. 0 23. 0 24. 0 25. 0 26. 5 27.0 27. 9 29. 0 30. 0 30. 0 262. 5 from the Zn 21. 0 23.0 24. 0 25. 0 26.5 27. 0 27. 0 29. 0 30. 0 30. 0 262. 5 21. 0 23. 0 24. 0 25. 0 26. 5 27.0 27. 0 29. 0 30. 0 30. 0 262. 5 21. D 23. 0 24. 0 25. 0 26. 5 18. 0 18.0 29. 0 30. 0 30. 0 244. 5 21. 0 23. 0 24.0 25. 0 25.0 18.0 9. 0 4. 0 4.5 4. 5 158.0 2i. 0 23. 0 24. 0 25. 0 26. 5 27. 0 27. 0 29. 0 30. 0 30. 0262. 5

21. 0 23. 0 24. 0 25. 0 26. 5 27. 0 29. 0 30. 0 32. 0 33. 0 270. 5 21.023. 0 24. 0 25. 0 26. 5 27. 0 29. 0 30. O 32. 0 33. 0 270. 5 21. D 23. 024. 0 25. 0 26. 5 27. 0 29. 0 30. 0 32. 0 33.0 270. 5 21. I) 23.0 24.025.0 26.5 9. 0 9.0 8. 0 12.0 12.0 169. 5 Control 21. 0 23. 0 24. 0 25.026. 5 27. 0 29.0 30. O 32. 0 33. 0 270. 5

(c) DEATHS, HEMAFEC-IA, NUMBER OF OOCYSTS IN FECES AND BOD Y WEIGHT GAINBody weight: Average per chicken. Judgment of hernafeoia (bloody partsin total feces evacuated at night): +=1 to 10, ++=11 to 20, +++=2l to30, ++++=31 to 40, +++++=more than 41. OPG: Number of oocysts in i g. offeces evacuated at night.

Days after in ection of oocysts Before infection 2 4 5 6 7 8 9 TotalPercent number body weight Age of chicken in days deaths gain (I) Whenthe number of oocysts infected is 0.3%, everyday:

Body weight (g.) g 0 51. 0 Death 0.3% from the 2nd day:

ody weight (g.). Hemafecia 0 49 2 O P G Death-.. 0.3%, from the 3rd day:

Body weight (g.) 51 s 01 o Death 0.3%, from the 4th day:

Body weight (g.) geprnaiecim 2 41. 1 Death n (C) DEATHS, HEMAFECIA,NUMBER OF OOCYSTS IN FECES AND BODY WEIGHT GAINCntinued Before Daysafter infection of oocysts infection 6 7 Total Percent Age of chicken indays number of deaths 0.2%, everyday:

Body weight (g.) Hemafeeia. OF G Death 0.1%, everyday:

Body weight (g.) Hernafecia OP G Death Infected and no administration:

Body weight (g.) glemafccia Control mninfected and no adminis- 1o Bodyweight (g.) Hemafecia OP G Death (II) When the number of oocystsinfected is 0.2%, everyday:

Body weight (g.) Hemafeeia OP G Death 0.2%, from the 2nd day:

Body weight (g.) Hemafeeia Death 0.2%, from the 3rd day:

Body weight (g.) Hernafecia OP G Death Infected and no administration:

Body weight (g.) Hemafecia OP G Deat Control atio Body weight (g.)Hemafecia O P G a Death h (uninfected and no adminis- (d) OOCYSTS INCAECUM In the caecum of all dead chickens except those infected with10x10 of oocysts and dead on the 7th day, on oocyst was observed. In thecaecum of the chickens dead within 7 days of the infected and notadministered group, 1,920,000 or 3,200,000 of oocysts were found. On the10th day after the start of the test, the survivals of all the groupswere sacrificed and subjected to the observation of oocysts. About110,000 of oocysts were found in the caecum of four survivals of thegroup infected with 8x10 of oocysts and not administered, and 500,000 inthat of two survivals of the group infected with 10 10 of oocysts andnot administered. Only about 1,000 oocysts were found in the caecum of 8birds whidh received everyday the drug at 0.1% concentration and nooocyst was found at all in other administered groups.

(e) VIEW OF POST-MORTEM EXAMINATION OF CAECUM The caecum of chickensdead within 5 to 6 days after infection (5 and 6 birds of the infectedand not ad ministered groups and 2 birds of the group administered with0.3% concentration from the 4th day) assumed a dark purple colour in itsentirety, showed hypertrophia and haemorrhagia and was filled with bloodclot. That of other dead chickens (each 1 bird dead on the 7th and 8thdays of the group administered with 0.1% concentration and 1 bird deadon the 7th day of the infected and not administered group) assumed adark purple colour, and was swollen 2 to 3 times and filled with bloodclot. That of 2 birds dead on the 7th day of the group infected with 1010 oocysts and not administered was swollen and filled with grayishwhile solid content.

On the contrary, that of the chickens of the groups respectivelyadministered with 0.2% concentration everyday and from the 2nd day, andwith 3% concentration everyday and from the 2nd day was normal showingno difference from that of the control group. The caecum of the chickensof the groups respectively administered with 0.1% concentrationeveryday, 0.2% concentration from the 3rd day and 0.3% concentrationfrom the 3rd and 4th days was slightly swollen and the grayish white toyellowish gray-white or reddish black content was present.

(5) T est on toxicity if drug White Leghorn male chickens aged 15 dayswere respectively given the mixtures of the compound in feed atconcentrations of 1%, 0.5%, 0.3% and 0.1% for 23 to 30 consecutive days.The result of the observation on the amount of feed taken and the bodyWeight gain in the said duration is shown in the subsequent table. Thegeneral symptons and the view of post-mortem examination are as follows:

white colour and the urinary tract had a small amount of uric acidcalculus.

Before Days of medication medication Age of chicken in days m no)unt offeed taken 20. 0 19.0 18.0 16. 0 18. 0 16. 5 10.0 *13. 0 16. 0 *11. O*20. 0 18. 0 *24. 0 24. 0 19. D 24. 0 20. 0 20. 0 20. 0 22. 0 20. 0 26.5 25.0 27.0 28. 5 31.0 25. 0 21.0 24. 0 36. 0 24. 0 *24. 0 20.0 21. 021. 0 24. 0 25.0 26. 5 27.0 29. 0 30. 0 31. 0 35.0 34. 0 36.0 36. 0 36.0 36.0 7 20. 0 21.0 21.0 24. 0 25. 0 26. 5 27.0 29. 0 30.0 31.0 35.0 36.0 36. 0 36.0 36.0 36.0 20. 0 21.0 21.0 24. 0 25.0 26. 5 27.0 29.0 30. 032. 0 35. O 36. 0 36. 0 36. 0 36. 0 36.0 Body weight Days of medication16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 Total amount Age of chickenin days AI(110;JI15 of feed taken Percent body weight gain Body weightgain (an):

NOTEs.-(1) The amount of feed taken and the body weight gain are shownby the average per test chicken (g.).

(2) The number in parentheses represents the amount of feed taken or thebody weight after the interruption of the medication.

(3) The total amount and the percent body (4) Asterisk represents deathof test chickens.

(0.) ACTIVITY OF CHICKENS The group which received the feed mixed withthe drug at 1% concentration gradually became melancholic, hanged downthe wings and behaved very inactively from the 3rd or 4th day after themedication. The administration in this group was interrupted on the 23rdday and, after the interruption, the inclination of recovery in activitywas observed.

The group which received the feed mixed with the drug at 0.5%concentration gradually became dull in activity from the 12th or 13thday after the administration and fell into melancholy.

Other administered groups showed almost no diiference from the controlgroup.

(b) VIE\V OF POST-MOR'I'EM EXAMINATION All dead chickens werecomplicated with fibrin-nus pericarditis, fibrinous pleurisy and lobarpneumonia. The kidney suffered from uratosis and assumed a grayish whitecolour. The urinary tract was choked with uric acid calculus to causeparacrisis of urine.

In 4 surviving birds of the group administered with the drug at 1%concentration, the liver was somewhat enlarged, the kidney fell inuratosis to assume a grayish weight gain show the values up to the 23rdday after the administration.

1. A method for preventing and treating poultry cocgenerally employedconcentration, for instance, lower than 0.3% and can be used verysafely.

An illustrative poultry feed composition useful for carrying out thepresent invention is, for instance, as follows:

Ingredient: Percent by weight Barley 18.5 Wheat 30 Rice 15 Bran 10Peanut meal 4 Soybean meal 5 Bone meal 1.3 Mineral salts, trace elementsvitamins, antibiotics 2 As aforeindicated, any conventional feed forpoultry can be used as carrier for the CS of this invention. It is 9 l0preferred to use any of the standard feed compositions a therapeuticallyeffective amount of 2-methyl-4-sulfanilrecommended by the AmericanNutrition Research Counamido-6-methoxy-3 (2H) pyridazinone. i1 2. Amethod according to claim 1, wherein the com- The composition isadministered as hereinbefore indi- Pound admlnlstefed to 4 y 01% to 02%Gated 5 (by weight) concentration in feed.

What is claimed is: References Cited by the Examiner Other administeredgroups showed no difference from Chem (London) 1948 page 2195 thecontrol group with no abnormality.

As illustrated above, the drug shows no toxicity at a SAM ROSEN PrimaryExaminercidiosis, which comprises orally administering to poultry 10 R.BARRESE, Assistant Examiner.

1. A METHOD FOR PREVENTING AND TREATING POULTRY COCCIDIOSIS, WHICH COMPRISES ORALLY ADMINISTERING TO POULTRY A THERAPEUTICALLY EFFECTIVE AMOUNT OF 2-METHYL-4-SULFANILAMIDO-6-METHOXY-3(2H)-PYRIDAZINONE. 